Figure 1a-Labeled LB Agar Ampicillin plate containing 25 micro-liters of organism BL21(DE3) made on 2/27/2014 by Justin and Xenia; plate was left overnight in incubator at 37 degrees Celsius; Label is incorrect in regards to presence of plasmid and ampicillin; No DNA control plate shows no colonies
Figure 1b- Labeled LB Agar Ampicillin plate containing 50 of micro-liters of organism BL21(DE3), micro-liters of plasmid pGEM-gbr22, and SOC media mixture; Made on 2/27/2014 by Justin and Xenia; plate was left in incubator overnight at 37 degrees Celsius; Experimental plate shows no colonies
Figure 1c- Agar plate with no antibiotic containing microorganisms from coughs; Plate was left overnight in incubator at 37 degrees Celsius; Made on 2/27/2014 by Justin and Xenia; plate shows no colonies
Figure 1d- Labeled LB Agar Ampicillin plate containing 50 micro-liters of organism BL21(DE3), plasmid pGEM-gbr22, and SOC media mixture; Made on 2/27/2014 by Ariana and Avery; Culture was grown overnight in incubator at 37 degrees Celsius; Experimental plate shows approximately 1200 colonies
Figure 2a- Labeled 125mL Erlenmeyer flask containing 0.625mL of starter culture (BL21(DE3) with pGEM-gbr22) mixed with 25mL LB and 50 micro-liters Ampicillin; made on 2/28/2014 by Justin; Culture was grown for 24 hours in shaking incubator at 37 degrees Celsius; Exhibits purple color
Figure 2b- Labeled 125mL Erlenmeyer flask containing 0.625mL of starter culture (BL21(DE3) with pGEM-gbr22) mixed with 25mL LB and 50 micro-liters Ampicillin; made on 2/28/2014 by Justin; Culture was grown for 24 hours in shaking incubator at 37 degrees Celsius; Exhibits purple color; Back-up sample
Figure 3a- Labeled 50mL conical tube containing pellet of culture (BL21(DE3) with pGEM-gbr22); Weight of pellet is 0.5 grams; Made on 3/1/2014 by Justin; Pellet exhibits purple color
Figure 3b- Labeled 50mL conical tube containing pellet of culture (BL21(DE3) with pGEM-gbr22); Weight of pellet is 0.4 grams; Made on 3/1/2014 by Justin; Pellet exhibits purple color
Figure 4a- Purified sample of purple protein, gbr22; sample has undergone one elution using the elution buffer of 1x PBS and 250mM imidazole during column chromatography; has volume of 5.0mL
Figure 5- Absorption spectrum using Nanodrop (A280 mode) of purified protein sample after one elution; absorption was measured at 280nm using 1cm path length; first trial; sample yielded 0.8495mg gbr22 using 280nm wavelength absorption
Gel electrophoresis of purple protein samples before drying: cell lysate, soluble fraction, flow through, wash, elution 1, elution 2, elution 1 (different person's protein), elution 2 (different person's protein); gel ladder was included; placed on Whatman paper
Dried gel electrophoresis of purple protein placed on Whatman Paper and wrapped in cellophane; wells are labeled with the first well being empty and the second being the ladder
Protein standards and ladders for gel electrophoresis
Introduction:
Materials & Methods:
Results
Discussion:
Conclusions:
References: